Cucurbita pepo hybrids

ABSTRACT

A plant or a part thereof, the plant being a  Cucurbita pepo  hybrid having more than one flower/fruit per node as in  Cucurbita pepo  subsp.  texana,  the fruit of the plant having the phenotype of that of  Cucurbita pepo  subsp.  pepo.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No. 13/389,608 filed on Feb. 9, 2012, which is a National Phase of PCT Patent Application No. PCT/IL2010/000644 having International Filing Date of Aug. 10, 2010, which claims the benefit of priority under 35 USC 119 (e) of U.S. Provisional Patent Application No. 61/272,029 filed on Aug. 10, 2009, the contents of which are incorporated herein by reference in their entirety.

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to a Cucurbita pepo hybrid having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo

The Cucurbita pepo L. is one of the most widely grown species of vegetable crops. It is the most familiar species of Cucurbita L., a genus native to the Americas which encompasses plants bearing edible fruits known as pumpkins and squash Like most other members of the Cucurbitaceae, Cucurbita plants are herbaceous, bearing large, palmate leaves and prominent fruits. Most species of Cucurbita are mesophytes, have fibrous root systems, and are monoecious, bearing large, intensely orange-yellow, nectar-producing, unisexual flowers that are foraged by bees. Each flower opens and is functional for one day and only during the early morning hours, withering by afternoon.

Cucurbita pepo is a collection of interfertile domesticated, feral, and wild plants. On the basis of allozyme variation and seed morphology, it has been classified into three subspecies, pepo, texana, and fraterna. Most cultivars belong to subsp. pepo but wild plants of this subspecies have not yet been found. The other cultivars belong to subsp. texana, which grows wild in central and southeastern U.S.A. Subsp. fraterna consists of wild specimens from northeastern Mexico, only.

Cucurbita pepo is perhaps the most polymorphic species in the plant kingdom. Its fruits range in size to over 20 kg; in shape from round to flat-scalloped, to long, bulbous cylindrical over 75 cm long; exterior color is based on hues of green, orange, and yellow, with color intensity ranging from pale to very intense, and gray contribution (darkness) ranging from none to very dark. Variegation, including striping and bicolor, can result in as many as four colors on the surface of the same fruit. Fruit mesocarp can be relatively thin or thick, and its color varies in the range from greenish white to white, yellow, light orange, and intense orange. Fruits rinds can be lignified or non-lignified, and smooth, warted, wrinkled, or netted.

Cucurbita pepo fruits are often used for culinary purposes when they are mature, 40 or more days past anthesis. However, the great economic value of this species rests on the common use of the young fruits, usually 2 to 5 days past anthesis, as food. These young fruits are known as summer squash. Summer squash are borne beginning approximately 50 days after seeding and as C. pepo grows well in a wide range of climates, it is very widespread in cultivation.

Most of the groups are centuries old. Some, the Pumpkin, the Acorn, and the Scallop, are indeed quite old, having been bred by native Americans prior to the

European contact at the end of the 15^(th) century. The Cocozelle and the Zucchini originated in southern and northern Italy, respectively. The Cocozelle is an old group, with records dating to the late 16^(th) century, and the Zucchini is the youngest group, with records dating only to the beginning of the 20^(th) century. The Cocozelle has some economic importance in Europe and in Israel yet, today, the Zucchini is by far the economically most important cultivar-group of Cucurbita pepo, perhaps exceeding in economic value the rest of the species, indeed, the rest of the genus combined.

Due to the high commercial value resting in the flowers and young fruits of Cucurbita pepo there is a need to develop new cultivars of this species having higher yields.

SUMMARY OF THE INVENTION

According to an aspect of some embodiments of the present invention there is provided a plant or a part thereof, the plant being a Cucurbita pepo hybrid having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

According to an aspect of some embodiments of the present invention there is provided a plant or a part thereof, the plant being a hybrid Cucurbita pepo subsp. pepo, having more than one flower/fruit per node at most nodes, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

According to an aspect of some embodiments of the present invention there is provided a plant or a part thereof of Cucurbita pepo selected from a population of progeny of the hybrid produced by crossing Cucurbita pepo subsp. texana and Cucurbita pepo subsp. pepo, the plant having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

According to an aspect of some embodiments of the present invention there is provided a plant or a part thereof, the plant being a Cucurbita pepo Zucchini hybrid, whose seeds have been deposited as Accession No. NCIMB 41744 (herein referred to as Multizuq).

According to an aspect of some embodiments of the present invention there is provided a plant or a part thereof, the plant being a Cucurbita pepo Cocozelle hybrid, whose seeds have been deposited as Accession No. NCIMB 41794 (herein referred to as Nizzan).

According to an aspect of some embodiments of the present invention there is provided a cell having the genome of the plant.

According to an aspect of some embodiments of the present invention there is provided a culture comprising a plurality of the cells.

According to some embodiments of the invention, the plant part is selected from the group consisting of roots, stems, leaves, cotyledons, flowers, fruit, embryos, pollen, and seeds.

According to some embodiments of the invention, the cell being regeneratable.

According to an aspect of some embodiments of the present invention there is provided a method of producing a Cucurbita pepo hybrid, the method comprising:

-   -   (a) crossing Cucurbita pepo subsp. texana with Cucurbita pepo         subsp. pepo;     -   (b) selecting progeny individuals from said crossing having more         than one flower/fruit per node as in Cucurbita pepo subsp.         texana, the fruit of said individuals having the phenotype of         that of Cucurbita pepo subsp. pepo.

According to some embodiments of the invention, the method further comprising:

-   -   (c) backcrossing individuals selected in step (b) with Cucurbita         pepo subsp. pepo.

According to some embodiments of the invention, said method additionally comprises the step of propagating said individuals following step (b) or (c).

According to some embodiments of the invention, the step of propagating includes the step of vegetative propagation.

According to some embodiments of the invention, the step of propagating includes the step of propagation by seed.

According to an aspect of some embodiments of the present invention there is provided a method for producing a Cucurbita pepo hybrid seed comprising crossing a first parent Cucurbita pepo plant with a second parent Cucurbita pepo plant and harvesting the resultant hybrid F₁ seed, wherein at least one of the first or the second parent Cucurbita pep. plant is the above plant.

According to an aspect of some embodiments of the present invention there is provided a hybrid seed produced by the above-method.

According to an aspect of some embodiments of the present invention there is provided a hybrid plant, or parts thereof, produced by growing the hybrid seed.

According to an aspect of some embodiments of the present invention there is provided a method of generating a Cucurbita pepo fruits and/or flowers, the method comprising:

-   -   (a) seeding seeds the above-hybrid and/or planting seedlings of         said seeds;     -   (b) growing plants generated from said seeds or said seedlings;         and     -   (c) harvesting the fruits and/or flowers of said plants, thereby         generating the Cucurbita pepo fruits and/or flowers.

Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying images. With specific reference now to the images, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the images makes apparent to those skilled in the art how embodiments of the invention may be practiced.

In the drawings:

FIG. 1 is a schematic representation of fruit shape of the eight edible cultivar-groups of Cucurbita pepo (after Paris, 1986, prior art figure).

FIG. 2 is an expanded schematic representation of fruit shape of the eight edible-cultivar-groups of Cucurbita pepo.

FIG. 3 is a photograph showing representative immature (2-5 days past anthesis) fruits of 23 cultivars Cucurbita pepo, at least one from each of the edible-fruited cultivar groups. Left to right, top row, Scallop Group: ‘Yellow Bush Scallop’, ‘Belye’, and “Sunburst'; Acorn group: ‘Royal Acorn’ and ‘Table Queen’; Crookneck Group: ‘Early Golden Crookneck’ and ‘Yellow Summer Crookneck’; Straightneck Group: ‘Early Prolific Straightneck’; center row, Pumpkin Group: ‘Ronde de Nice’ and ‘Tondo Scuro di Piancenza’; Vegetable Marrow Group: ‘Verte Petite d'Alger’ (syn. ‘Grey Zucchini’), ‘All Green Bush’, ‘Beirut’, ‘Long Green’, and ‘Sihi Lavan’; bottom row, Cocozelle Group: ‘Romanesco’, ‘Lungo di Toscana’ (with corolla and calyx), ‘Long Cocozelle’, ‘Striato d'Italia’, and ‘Lungo Bianco di Sicilia’; Zucchini Group: ‘Black Zucchini’, ‘Nano Verde di Milano’, and ‘True French’.

FIG. 4 is a photograph of a plant of ‘Multizuq’ Zucchini hybrid. Note the Zucchini-type fruit and the presence of more that one flower bud per leaf axil.

FIG. 5 is photograph showing fruits of ‘Multizuq’ Zucchini squash hybrid.

FIG. 6 is a photograph showing a plant of ‘Nizzan’ Cocozelle hybrid. Note the presence of more than one flower bud per leaf axil.

FIG. 7 is a photograph showing fruits of ‘Nizzan’ Cocozelle squash hybrid.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention, in some embodiments thereof, relates to a Cucurbita pepo hybrid having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

Vegetables of the Cucurbita pepo species are of significant economic importance as they encompass many coveted summer and winter edible types as well as ornamental gourds. Cucurbita pepo refers to what are commonly known as the summer squash, that is, cocozelle, vegetable marrow, scallop, zucchini, straightneck and crookneck types and winter squash such as acorn and pumpkin.

Accordingly, the inventor of the present invention has evaluated options for increasing fruit/flower yield of Cucurbita pepo.

The inventor noticed that there is a difference in the number of flowers borne in and next to the leaf axils in Cucurbita pepo: in C. pepo subsp. pepo, only one flower bud is formed in or next to each leaf axil. However, in C. pepo subsp. texana, more than one flower can be borne in or next to the leaf axil in each of its four edible-fruited cultivar-groups. This characteristic is most strongly expressed in the Crookneck Group, in which three or even more flowers can be found in or next to the leaf axils. When two or more female flowers are produced, plants of Crookneck cultivars can produce more than one fruit at the same node. Some Crookneck cultivars, such as ‘Supersett’, are known to be extremely high yielding.

The present invention is of Cucurbita pepo hybrids having more than one flower/fruit per node (as in Cucurbita pepo subsp. texana), but with the appearance of the fruit of Cucurbita pepo subsp. pepo.

This invention was demonstrated by the development Zucchini and Cocozelle hybrids having the multiple-flowering characteristic. This was accomplished by plant breeding methodology, specifically, the backcross-pedigree selection method (Allard, 1960). As the donor of the multiple-flowering characteristic, the Crookneck, is so distant in genealogy from the Zucchini and the Cocozelle, this became a long and tedious process, but unexpectedly and surprisingly appears to have been successful.

Thus, according to an aspect of the invention there is provided a plant or a part thereof, the plant being a Cucurbita pepo hybrid having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

According to an aspect of the invention there is provided a plant or a part thereof, the plant being a hybrid Cucurbita pepo subsp. pepo, having more than one flower/fruit per node at most nodes (i.e., more than 50% of the nodes), the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

According to an aspect of the invention there is provided a plant or a part thereof of Cucurbita pepo selected from a population of progeny of the hybrid produced by crossing Cucurbita pepo subsp. texana and Cucurbita pepo subsp. pepo, the plant having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

The phrase “plant or part thereof” as used herein encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, fruits, flowers, shoots, stems, roots (including tubers), pollen, embryo, cotyledons, leaves and plant cells, tissues and organs. The plant may be in any form including suspension cultures, embryos, meristematic regions, callus tissue, leaves, gametophytes, sporophytes, pollen, ovules and microspores.

As used herein “Cucurbita pepo” refers to the collection of interfertile domesticated, feral and wild plants of the subspecies pepo, texana, and fraterna.

As used herein the term “hybrid” refers to the offspring derived from crossing two parental breeding lines of Cucurbita pepo.

The hybrid is heterozygous and derived from the crossing of the two parental breeding lines, each of which is nearly homozygous. The hybrid and each of its two parent breeding lines are homogeneous populations. The hybrid and its parents according to the invention contain, however, at least a small portion of the genome, by introgression, of Cucurbita pepo subsp. texana, specifically, the gene or genes conferring the production of more than one flower/fruit per node and adjacent chromosomal regions.

On the basis of fruit shape, Cucurbita pepo is considered to consist of eight edible-fruited cultivar-groups (FIGS. 1, 2, and 3), four of which are classified in C. pepo subsp. pepo and the other four in C. pepo subsp. texana. Description of the eight groups and their placement in the two cultivated subspecies are summarized in Table 1, below. The fruits of two of the cultivar-groups, Pumpkin and Acorn, are used primarily when mature. Pumpkin fruits are round, being spherical, oblate, globular, or oval and Acorn fruits are turbinate with alternating longitudinal ridges and furrows. In both of these groups, the length-to-width ratio of the fruits is approximately 1:1. The fruits of the other six cultivar-groups are used when immature, as summer squash, and diverge markedly from this 1:1 ratio. Scallop squash are flat and scalloped, hence having a length-to-width ratio that is considerably less than 1:1. The other five groups, Cocozelle, Crookneck, Straightneck, Vegetable Marrow, and Zucchini, have a length-to-width ratio that is considerably greater than 1:1 (Table 1, below). Each cultivar-group of both subspecies is comprised of numerous cultivars. The name of one representative cultivar of each cultivar-group is given in Table 1.

Plants of the invention have more than one flower/fruit per node. Flower buds in Cucurbita pepo are differentiated in or beside the leaf axils, that is, at the junctions of the petiole bases with the stem; these junctions are also referred to as stem nodes. Thus, according to embodiments of the invention, 2, 3, 4 or even more flower buds can be formed at each stem node.

As mentioned, plants of the invention produce more than one flower at most stem nodes and have fruits which are endowed with the phenotype of Cucurbita pepo subsp. pepo. The potentially higher yield of flowers and fruits per hybrid plant as compared to all previously existing Cucurbita pepo subsp. pepo does not compromise fruit phenotype.

As used herein “fruit phenotype” refers to the fruit shape, specifically, fruit profile and topography. Table 1 below lists the fruit shape profiles of the cultivar-groups belonging to Cucurbita pepo subsp. pepo and C. pepo subsp. texana. The fruits of C. pepo subsp. pepo can be differentiated from those of C. pepo subsp. texana in fruit topography. The fruits of the Cocozelle Group are nearly smooth to obviously ribbed along their longitudinal axis. The Zucchini has uniformly cylindrical fruits which are slightly ribbed along their longitudinal axis. These ribs are promonitories occurring above the 10 main vascular tracts running along the longitudinal axis of the fruits. In contrast, the fruits of the cultivar-groups of C. pepo subsp. texana, have furrows along their longitudinal axis. These furrows are depressions occurring above the 10 main vascular tracts. Both, ribs and furrows, are easily observed in cross-cut sections of the fruits.

TABLE 1 The edible-fruited cultivar-groups of Cucurbita pepo (after Paris, 2000). Cultivar- group Synonyms Subspecies Standard cultivar Fruit shape Pumpkin pepo Connecticut Field Round: spherical oblate, ovate, globular Vegetable Middle pepo Beirut Short, tapered cylindrical, narrow at peduncle Marrow eastern end, broad at stylar end, length-to-broadest width ratio ranging from 1.5-3.0 Cocozelle Italian pepo Striato d'Italia Long to extremely long, cylindrical, bulbous near stylar end, length-to-broadest width ratio at least 3.5 Zucchini Courgette pepo Black Zucchini Uniformly cylindrical, length-to-broadest width ratio 3.5-5.0 Acorn Table texana Table Queen Turbinate, furrowed, broad at peduncle end, Queen convex at stylar end Crookneck texana Yellow Summer Elongated, with narrow, slightly to very curved Crookneck neck, broad stylar half, convex stylar end Scallop Patty Pan, texana Golden Bush Flattened, with scalloped margins Patisson Scallop Straightneck texana Early Prolific Cylindrical, with short neck or constriction near Straightneck the stem end and broad stylar half, convex or pointed distal end

According to one embodiment of the invention the hybrid plant is a Zucchini and crookneck hybrid.

One specific embodiment of the invention is the hybrid ‘Multizuq’, which is the F1 hybrid of zucchini breeding line 1688-1-3-16 crossed with zucchini breeding line 1477-1-7-2-10. Seeds of ‘Multizuq’ were deposited under the Budapest treaty on Jul. 30, 2010 at the NCIMB Ltd. Scotland UK, under the accession number NCIMB 41744.

According to one embodiment of the invention the hybrid plant is a cocozelle and crookneck hybrid.

One other specific embodiment of the invention is the hybrid ‘Nizzan’, which is the F1 hybrid of cocozelle breeding line 1260-4-6-2-10 crossed with cocozelle breeding line 1413-4-54-7. Seeds of Nizzan were deposited under the Budapest treaty on Dec. 10, 2010 at the NCIMB Ltd. Scotland UK, under the accession number 41794.

The invention also relates to a cell having the genome of the plant of the invention i.e., having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo. According to a specific embodiment the cells are regenerable.

The invention also relates to a culture comprising a plurality of the cells.

According to an embodiment of the invention the plant is selected from a population of progeny of the hybrid produced by crossing within Cucurbita pepo subsp. pepo, the plant having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo.

Thus according to an aspect of the invention there is provided a method of producing a Cucurbita pepo hybrid, the method comprising:

-   (a) crossing Cucurbita pepo subsp. texana with Cucurbita pepo subsp.     pepo; -   (b) selecting a progeny individuals from said crossing having more     than one flower/fruit per node as in Cucurbita pepo subsp. texana,     the fruit of said individuals having the phenotype of that of     Cucurbita pepo subsp. pepo.

According to a specific embodiment, the method further comprising:

(c) backcrossing individuals selected in step (b) with Cucurbita pepo subsp. pepo.

The term “backcrossing” as used herein refers to the repeated crossing back to elite breeding lines of C. pepo subsp. pepo. The parental plant which contributes the desired characteristic is termed the donor parent. This terminology refers to the fact that the donor parent is used once, in the initial cross. The parental line to which the special characteristic from the donor parent is introgressed is known as the recurrent parent, as it is used repeatedly (recurrently) in the backcrossing protocol.

In a typical backcross protocol, a plant from the original line of interest (recurrent parent) is crossed with a plant from the second variety, the donor parent, which carries the single special characteristic of interest. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a plant is obtained wherein essentially all of the desired morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred (introgressed) characteristic from the donor parent.

Backcrossing methods can be used with the present invention to improve or introduce a unique characteristic into the parent lines.

According to a specific embodiment the method additionally comprises the step of propagating said individuals following step (b) or (c).

According to a specific embodiment the method the step of propagating includes the step of vegetative propagation.

According to a specific embodiment the method the step of propagating includes the step of propagation by seed.

The development of hybrids requires, in general, the development of homozygous inbred lines, the crossing of these lines, and the evaluation of the crosses. Pedigree breeding and recurrent selection breeding methods are used to develop inbred lines from breeding populations. Breeding programs combine the genetic backgrounds from two or more inbred lines or various other germplasm sources into breeding pools from which new inbred lines are developed by selfing and selection of desired phenotypes. The new inbreds are crossed with other inbred lines and the hybrids from these crosses are evaluated to determine which of those have commercial potential.

Pedigree breeding starts with the crossing of two genotypes, each of which may have one or more desirable characteristics that is lacking in the other or which complements the other. If the two original parents do not provide all the desired characteristics, other sources can be included in the breeding population. In the pedigree method, superior plants are selfed and selected in successive generations. In the succeeding generations the heterozygous condition gives way to homogeneous lines as a result of self-pollination and selection. Typically in the pedigree method of breeding five or more generations of selfing and selection is practiced: F1 to F2; F3 to F4; F4 to F5, etc.

A single cross hybrid results from the cross of two inbred lines, each of which has a genotype that complements the genotype of the other. The hybrid progeny of the first generation is designated F1. In the development of commercial hybrids only the F1 hybrid plants are sought. Preferred F1 hybrids are more vigorous than their inbred parents. This hybrid vigor, or heterosis, can be manifested in many polygenic traits, including increased vegetative growth and increased yield

Cucurbita pepo plants can be easily cross-pollinated. A trait is readily transferred from one plant to another plant, including Cucurbita pepo plants of different subspecies, using conventional breeding techniques, for example to obtain new commercializable hybrids. The introgression of a trait into the elite line is for example achieved by recurrent selection breeding, for example by backcrossing. In this case, the elite line (recurrent parent) is first crossed to a donor inbred (the non-recurrent parent) that carries the trait. The progeny of this cross is then mated back to the recurrent parent followed by selection in the resultant progeny for the trait. After three, preferably four, more preferably five or more generations of backcrosses with the recurrent parent with selection for the trait, the progeny is heterozygous for the locus harboring the resistance, but is like the recurrent parent for most or almost all other genes (see, for example, Poehlman & Sleper (1995) Breeding Field Crops, 4th Ed., 172-175; Fehr (1987) Principles of Cultivar Development, Vol. 1: Theory and Technique, 360-376, incorporated herein by reference). Selection for the trait is carried out after each cross.

In one embodiment, a plant of the present invention is an inbred, a hybrid, or a dihaploid, for example produced by pedigree breeding or by recurrent selection breeding. In one embodiment, a plant of the present invention has commercially acceptable horticultural characteristics.

In one embodiment, the present invention discloses a method of producing seed of a plant of the present invention (i.e., having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo) comprising: a) growing a plant of the invention; b) allowing said plant to self-pollinate; c) harvesting seeds from said plant. As mentioned plants can also be propagated vegetatively using methods well-known in the art, for example in-vitro plant tissue culture, rooting side shoot or protoplast fusion. In one embodiment, a method of vegetatively propagating a plant of the present invention comprises: a) collecting tissue of a plant of the present invention; b) cultivating said tissue to obtain proliferated shoots; c) rooting said proliferated shoots to obtain rooted plantlets; d) growing plants from said rooted plantlets; and harvesting seeds from said plants.

Plants of the invention (i.e., having more than one flower/fruit per node as in Cucurbita pepo subsp. texana, the fruit of the plant having the phenotype of that of Cucurbita pepo subsp. pepo) can also be transformed genetically with a gene of interest, using techniques well known in the art. Accordingly, the present invention also further discloses a Cucurbita pepo plant according to the instant invention, such as a gene of interest.

In one embodiment, the present application discloses methods of producing a fruit/flower, comprising growing a plant of the invention until a fruit or flower is produced and harvesting the fruit or flower. It will be appreciated that although the instant specification places an emphasis on fruit yield in Cucurbita pepo, the flowers have a culinary as well as ornamental value and therefore the present teachings are meant to encompass both fruit and flower products.

As used herein the term “about” refers to ±10%.

The terms “comprises”, “comprising”, “includes”, “including”, “having” and their conjugates mean “including but not limited to”.

The term “consisting” of means “including and limited to”.

The term “consisting essentially of” means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

As used herein, the singular form “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a compound” or “at least one compound” may include a plurality of compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases “ranging/ranges between” a first indicate number and a second indicate number and “ranging/ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

As used herein the term “method” refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non limiting fashion.

Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, “Molecular Cloning: A laboratory Manual” Sambrook et al., (1989); “Current Protocols in Molecular Biology” Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., “Current Protocols in Molecular Biology”, John Wiley and Sons, Baltimore, Ma. (1989); Perbal, “A Practical Guide to Molecular Cloning”, John Wiley & Sons, New York (1988); Watson et al., “Recombinant DNA”, Scientific American Books, New York; Birren et al. (eds) “Genome Analysis: A Laboratory Manual Series”, Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; “Cell Biology: A Laboratory Handbook”, Volumes I-III Cellis, J. E., ed. (1994); “Culture of Animal Cells—A Manual of Basic Technique” by Freshney, Wiley-Liss, N. Y. (1994), Third Edition; “Current Protocols in Immunology” Volumes I-III Coligan J. E., ed. (1994); Stites et al. (eds), “Basic and Clinical Immunology” (8th Edition), Appleton & Lange, Norwalk, Ct. (1994); Mishell and Shiigi (eds), “Selected Methods in Cellular Immunology”, W. H. Freeman and Co., New York (1980); available immunoassays are extensively described in the patent and scientific literature, see, for example, U.S. Pat. Nos. 3,791,932; 3,839,153; 3,850,752; 3,850,578; 3,853,987; 3,867,517; 3,879,262; 3,901,654; 3,935,074; 3,984,533; 3,996,345; 4,034,074; 4,098,876; 4,879,219; 5,011,771 and 5,281,521; “Oligonucleotide Synthesis” Gait, M. J., ed. (1984); “Nucleic Acid Hybridization” Hames, B. D., and Higgins S. J., eds. (1985); “Transcription and Translation” Hames, B. D., and Higgins S. J., eds. (1984); “Animal Cell Culture” Freshney, R. I., ed. (1986); “Immobilized Cells and Enzymes” IRL Press, (1986); “A Practical Guide to Molecular Cloning” Perbal, B., (1984) and “Methods in Enzymology” Vol. 1-317, Academic Press; “PCR Protocols: A Guide To Methods And Applications”, Academic Press, San Diego, Calif. (1990); Marshak et al., “Strategies for Protein Purification and Characterization—A Laboratory Course Manual” CSHL Press (1996); all of which are incorporated by reference as if fully set forth herein. Other general references are provided throughout this document. The procedures therein are believed to be well known in the art and are provided for the convenience of the reader. All the information contained therein is incorporated herein by reference.

Materials and Methods

Multiple flowering in Zucchini was realized in the development of the F₁ hybrid ‘Multizuq’ and its parents. These developments were the outcome of a complex series of crossing. The initial cross, designated No. 755, was made at the A.R.O. Newe Ya'ar Research Center, northern Israel. The female parent, the parent having the multiple-flowering characteristic, was the Crookneck hybrid ‘Supersett’ and the male parent was the open-pollinated Cocozelle ‘Striato Pugliese’. Seeds of ‘Supersett’ had been from Joseph Harris Co., Rochester, N.Y. and seeds of ‘Striato Pugliese’ had been obtained from Emanuele Larosa, Bari, Italy. F₁ plants were self-pollinated and an F₂ plant having the multiple-flowering characteristic of ‘Supersett’ was then crossed with an inbred of the open-pollinated Zucchini ‘True French’. Seeds of ‘True French’ had been obtained from Thompson & Morgan, U.K. F₁ plants were self-pollinated and some F₂ plants having multiple flowering were obtained. From this point on, a divergence into two separate parental lines was undertaken.

In the female parent line of ‘Multizuq’, a cross was made with a Zucchini inbred developed at Newe Ya'ar, herein designated 1-16. The F₁ was selfed, again F₂s were grown out and selected for multiple flowering, and this time were crossed on a different Zucchini inbred developed at Newe Ya'ar, herein designated 16-33. This was followed by four more cycles of backcrossing and selfing to the same, 16-33, inbred, following by three of generations of selfing to give the female parent of ‘Multizuq’, designated 1688-1-3-16.

In the male parent line of ‘Multizuq’, a multiple-flowering F₂ plant was backcrossed on ‘True French’. A new F₂ was obtained and a multiple-flowering individual was crossed on inbred 1-16. Once again, a self-pollination was made and a multiple-flowering F₂ plant was crossed again with inbred 1-16. After self-pollination, a multiple-flowering F₂ plant was crossed with a double-cross hybrid of complex parentage. Finally, this was crossed again with 1-16, and an individual in the F₂ selected for multiple flowering. This was followed by four more generations of selfing to give the male parent of ‘Multizuq’, designated 1477-1-7-2-10. ‘Multizuq’, therefore, is the F₁ hybrid of 1688-1-3-16 crossed with 1477-1-7-2-10.

Multiple flowering in Cocozelle was realized in the development of the F₁ hybrid ‘Nizzan’ and its parents. These developments were the outcome of a complex series of crossing. The initial crosses were the same as in the development of ‘Multizuq’, through the stage of selecting a multiple-flowering individual in the F₂ of the cross with inbred 1-16. In the female parent line of ‘Nizzan’, this F₂ multiple flowering individual was crossed with a Cocozelle inbred developed at Newe Ya'ar, herein designated 1-8-13. This was followed by four of generations of selfing to give the female parent of ‘Nizzan’, designated 1260-4-6-2-10.

In the male parent line of ‘Nizzan’, a multiple-flowering plant of the same F₂ was crossed on an inbred, designated 1-7, that was derived from a complex cross consisting mostly of Cocozelle parentage. A self-pollination was made on this F₁ and a multiple-flowering plant in the resulting F₂ was crossed with a commercially available Cocozelle F₁ hybrid. After self-pollination of a plant of this cross, a multiple-flowering F₂ plant was selected and self-pollinated. Self-pollination was repeated for several more generations until the true-breeding, multiple-flowering Cocozelle inbred designated 1413-4-54-7 was derived, which serves as the male parent of ‘Nizzan’.

‘Nizzan’, therefore, is the F₁ hybrid of 1260-4-6-2-10 crossed with 1413-4-54-7.

Results

‘Multizuq’ is a multiple-flowering hybrid Zucchini. It has open, non-branching, relatively small plants. The petioles are fairly smooth and the laminae are silver-mottled. More than one flower bud is formed in or beside the leaf axils, that is, at the junctions of the petiole bases with the stem (FIG. 4). The flower buds are usually not of the same size and do not reach anthesis on the same day. One or more of these buds may be male and one or more may be female. The fruits are typically Zucchini by being uniformly cylindrical of length-to-width ratio approximating 4:1 (Table 1). When young, at the summer squash age (2-5 days past anthesis), they are glossy and very intensely green (FIG. 5). The stylar scar is fairly small and the peduncle, while relatively short and thick, detaches easily from the stem when the fruit is picked. This hybrid is expected to realize its greatest potential in yield when grown under long-season, greenhouse conditions.

‘Nizzan’ is a multiple-flowering hybrid Cocozelle. It has open, non-branching, relatively small plants. The petioles are fairly smooth and the laminae are silver-mottled. More than one flower bud is formed in or beside the leaf axils, that is, at the junctions of the petiole bases with the stem (FIG. 6). The flower buds are usually not of the same size and do not reach anthesis on the same day. One or more of these buds may be male and one or more may be female. The fruits are typically Cocozelle by being long and bulbously cylindrical, of length-to-broadest-width ratio approximating 4:1 (Table 1). When young, at the summer squash age (2-5 days past anthesis), they are glossy and light green (FIG. 7). The stylar scar is fairly small and the peduncle is relatively long and narrow, detaching easily from the stem when the fruit is picked. This hybrid is expected to realize its greatest potential in yield when grown under long-season, greenhouse conditions.

Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

Literature Cited Other References are Listed Throughout the Specification

Allard, R. W. 1960. Principles of plant breeding, pp. 43-49. Wiley, N.Y.

Decker, D. S. 1985. Numerical analyzsis of allozyme variation in Cucurbita pepo. Econ. Bot. 39:300-309.

Decker, D. S. 1988. Origin(s), evoluation, and systematics of Cucurbita pepo. Econ. Bot. 42: 4-15.

Duchesne, A. N. 1786. Essai sur l'histoire naturelle des courges. Panckoucke, Paris.

Ferriol, M., B. Pico, and F. Nuez. 2003. Genetic diversity of a germplasm collection of Cucurbita pepo using SRAP and AFLP markers. Theor. Appl. Genet. 107: 271-282.

Paris, H. S. 1986. A proposed subspecific classification for Cucurbita pepo. Phytologia 61: 133-138.

Paris, H. S. 1989. Historical records, origins, and development of the edible cultivar groups of Cucurbita pepo (Cucurbitaceae). Econ. Bot. 43: 423-443.

Paris, H. S. 2000. History of the cultivar-groups of Cucurbita pepo, pp. 71-170, 4 pl., in J. Janick, editor, Horticultural Reviews 25(2001). Wiley, N.Y.

Paris, H. S. 2007. The drawings of Antoine Nicolas Duchesne for his natural history of the gourds. Museum National d'Histoire Naturelle, Paris.

Paris, H. S. 2008. Summer squash, pp. 351-379, in: J. Prohens and F. Nuez, editors, Handbook of Plant Breeding, Vegetables I. Springer, N.Y.

Paris, H. S., N. Yonash, V. Portnoy, N. Mozes-Daube, G. Tzuri, and N. Katzir. 2003. Assessment of genetic relationships in Cucurbita pepo (Cucurbitaceae) using AFLP, ISSR, and SSR markers. Theor. Appl. Genet. 106: 971-978. 

What is claimed is:
 1. A plant of Cucurbita pepo comprising a genome of Cucurbita pepo subsp. pepo plant of a cultivar group selected from the group consisting of Zucchini and Cocozelle and an introgression from Cucurbita pepo subsp. texana subsp. Crookneck, the plant having more than one flower/fruit per node as in Cucurbita pepo subsp. texana subsp. Crookneck conferred by said introgression, the fruit of the plant having a fruit shape of the cultivar group selected from the group consisting of Zucchini and Cocozelle, wherein the plant is obtainable by: (a) crossing a Cucurbita pepo subsp. texana subsp. Crookneck plant with a Cucurbita pepo subsp. pepo plant of a cultivar group selected from the group consisting of Zucchini and Cocozelle; (b) self-pollinating progeny individuals from said crossing; (c) selecting progeny individuals from said selfing having more than one flower/fruit per node as in Cucurbita pepo subsp. texana; and (d) backcrossing individuals selected in step (c) to a Cucurbita pepo subsp. pepo of a cultivar group selected from the group consisting of Zucchini and Cocozelle, wherein said steps (b) to (d) and optionally selfing following (d) is repeated for a sufficient number of times so as to obtain a plant having more than one flower/fruit per node as in Cucurbita pepo subsp. texana subsp. Crookneck, the fruit of the plant having the fruit shape of the cultivar group selected from the group consisting of Zucchini and Cocozelle.
 2. The plant of claim 1, being an inbred.
 3. A cell having the genome of the plant of claim
 1. 4. A culture comprising a plurality of the cells of claim
 3. 5. A plant part of the plant part of claim 1, selected from the group consisting of roots, stems, leaves, cotyledons, flowers, fruit, embryos, pollen, and seeds.
 6. The cell of claim 3, being regeneratable.
 7. A method of producing the plant of claim 1, the method comprising: (a) crossing a Cucurbita pepo subsp. texana subsp. Crookneck plant with a Cucurbita pepo subsp. pepo plant of a cultivar group selected from the group consisting of Zucchini and Cocozelle; (b) self-pollinating progeny individuals from said crossing; (c) selecting progeny individuals from said selfing having more than one flower/fruit per node as in Cucurbita pepo subsp. texana; and (d) backcrossing individuals selected in step (c) to a Cucurbita pepo subsp. pepo of a cultivar group selected from the group consisting of Zucchini and Cocozelle, wherein said steps (b) to (d) and optionally selfing following (d) is repeated for a sufficient number of times so as to obtain a plant having more than one flower/fruit per node as in Cucurbita pepo subsp. texana subsp. Crookneck, the fruit of the plant having the fruit shape of the cultivar group selected from the group consisting of Zucchini and Cocozelle.
 8. The method of claim 7, wherein said method additionally comprises the step of propagating individuals obtained following step (d).
 9. The method of claim 8, wherein the step of propagating includes the step of vegetative propagation.
 10. The method of claim 8, wherein the step of propagating includes the step of propagation by seed.
 11. A method for producing a Cucurbita pepo hybrid seed, the method comprising crossing a first parent inbred Cucurbita pepo plant with a second parent inbred Cucurbita pepo plant and harvesting the resultant hybrid F1 seed, wherein both parents are Cucurbita pepo plants of claim
 1. 12. A hybrid seed obtainable by crossing a first parent inbred Cucurbita pepo plant with a second parent inbred Cucurbita pepo plant, wherein both of the first and the second parent Cucurbita pepo plant are of the plant of claim
 2. 13. A hybrid plant, or parts thereof, produced by growing the hybrid seed of claim
 12. 14. A method of generating a Cucurbita pepo fruits and/or flowers, the method comprising: (a) seeding seeds of the plant of claim 1, and/or planting seedlings of said seeds; (b) growing plants generated from said seeds or said seedlings; and (c) harvesting the fruits and/or flowers of said plants, thereby generating the Cucurbita pepo fruits and/or flowers. 